The main abbreviation of NASBA is Nucleic Acid Sequence Based Amplification. This is a method in molecular biology. This amplification is used to amplify RNA sequences. In 1991, J Campton was developed this method NASBA. He defined that, “a premier-dependent technology that can be used for the continuous amplification of nucleic acids in a single mixture at one temperature.” This amplification was used for the rapid diagnosis and quantification of HIV-1 in patient sera after the invention of NASBA.
It works at isothermic conditions only (usually at a constant temperature of 41°C. This wonderful technique is used to improve rapid diagnostic tests for several pathogenic viruses with single stranded RNA genomes. For example: influenza A, foot and mouth disease virus, severe acute respiratory syndrome (SARS)-associated corona virus, and Human Boca virus (HBoV).
NASBA work as follows:
- RNA template is given to the reaction mixture, the first primer attaches to its complementary site at the 3′ end of the template.
- Reverse transcriptase synthesizes the opposite, complementary DNA strand.
- RNAse H destroys the RNA template (RNAse H only destroys RNA-DNA hybrids, but not single-stranded RNA)
- The second primer attaches to the 5′ end of the DNA strand.
- T7 RNA polymerase produces a complementary RNA strand which can be used again in step 1, so this reaction is cyclic.
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